Fast LC–MS/MS analysis of free oxysterols derived from reactive oxygen species in human plasma and carotid plaque

Helmschrodt, Christin; Becker, Stefan; Schröter, Jenny; Hecht, Max; Aust, Gabriela; Thiery, Joachim; Ceglarek, Uta

doi:10.1016/j.cca.2013.06.022

Cited by 68 publications

(46 citation statements)

References 32 publications

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“…This provided lipid classes; exact mass studies ( 27,28 ). The lipid ion peak at m/z 383.3 has been identifi ed to be dehydrated 7-ketocholestrol in a number of published studies involving human blood samples (29)(30)(31); although, we recognize that oxidation of cholesterol at some other position is possible and would result in similar fragmentation.…”

Section: Chemical Characterization Of the Validated Serum Lipid Preecmentioning

confidence: 94%

Detection and confirmation of serum lipid biomarkers for preeclampsia using direct infusion mass spectrometry

Anand

Young

Esplin

et al. 2016

Journal of Lipid Research

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Section: Chemical Characterization Of the Validated Serum Lipid Preecmentioning

confidence: 94%

Detection and confirmation of serum lipid biomarkers for preeclampsia using direct infusion mass spectrometry

Anand

Young

Esplin

et al. 2016

Journal of Lipid Research

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“…The method described by Helmschrodt (2013) was adopted for extraction of oxysterols from plasma. Briefly, thawed plasma samples were centrifuged at 3,200 × g for 5 minutes and 80 µl of plasma was added to vials containing 80 µl of a methanolic solution of a mixture of internal standards (400 pg of the deuterated version of each oxysterol) or blank (methanol without oxysterols).…”

Section: Methodsmentioning

confidence: 99%

“…On the day of analysis, extracts were reconstituted with 80 µl of a methanol/water mixture (3:1, v/v) and centrifuged prior to analysis for 5 min at 3,220 × g . All solutions contained 0.05% (v/v) butylated hydroxytoluene to prevent autoxidation (Helmschrodt 2013). …”

Section: Methodsmentioning

confidence: 99%

Selective changes in cholesterol metabolite levels in plasma of breast cancer patients after tumor removal

Souček

Vrána

Ueng

et al. 2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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Introduction Oxygenated metabolites of cholesterol (“oxysterols”) can influence carcinogenesis and contribute to resistance to endocrine therapy, an effect mostly described in vitro. Objectives We sought to establish a method for screening plasma levels of oxysterols in breast cancer patients, estimate their individual variability and detection limits, and provide basic information about their roles in tumor biology. Method Liquid-chromatography coupled with tandem mass spectrometry was used for determination of levels of 25-hydroxycholesterol, 27-hydroxycholesterol, 7α-hydroxycholesterol, and 7-ketocholesterol in plasma sample pairs from patients before and 12–24 months after surgical removal of tumors (n=24). Deuterated standards of all oxysterols were used for method validation. Result All oxysterols were successfully detected in patient plasma samples. A significant increase in the level of 7-ketocholesterol was observed in the samples following tumor removal and the start of therapy compared to the sampling before (p=0.002). This increase was unrelated to personal characteristics of patients, expression of estrogen receptor, or to adjuvant therapy type. Conclusion This study shows, for the first time, that circulating levels of oxysterols, especially 7-ketocholesterol, may reflect the presence of tumor cells in patients.

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“…Oxidative stress is intimately linked to the pathogenesis of vascular diseases such as atherosclerosis, diabetes, and hypertension [27, 28]. In the vascular wall, many enzymatic systems, including NADPH oxidase, produce reactive oxygen species (ROS) and contribute to vascular complications such as vascular hypertrophy and inflammation [27].…”

Section: Introductionmentioning

confidence: 99%

“…In the vascular wall, many enzymatic systems, including NADPH oxidase, produce reactive oxygen species (ROS) and contribute to vascular complications such as vascular hypertrophy and inflammation [27]. In fact, ROS levels are elevated in atherosclerotic plaques [28], suggesting a critical role for ROS in vascular disease [27, 28]. However, the detailed mechanisms involved in cellular adaptation to ROS produced by vascular cells remain to be elucidated.…”

Section: Introductionmentioning

confidence: 99%

Ligand-Activated Peroxisome Proliferator-Activated Receptor δ Attenuates Vascular Oxidative Stress by Inhibiting Thrombospondin-1 Expression

et al. 2018

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Thrombospondin-1 (TSP-1) is implicated in vascular diseases associated with oxidative stress, such as abdominal aortic aneurysms, ischemia-reperfusion injury, and atherosclerosis. However, the regulatory mechanisms underlying TSP-1 expression are not fully elucidated. In this study, we found that peroxisome proliferator-activated receptor δ (PPARδ) inhibited oxidative stress-induced TSP-1 expression and migration in vascular smooth muscle cells (VSMCs). Activation of PPARδ by GW501516, a specific ligand for PPARδ, significantly attenuated hydrogen peroxide (H₂O₂)-induced expression of TSP-1 in VSMCs. Small interfering RNA-mediated knockdown of PPARδ and treatment with GSK0660, a selective PPARδ antagonist, reversed the effect of GW501516 on H₂O₂-induced expression of TSP-1, suggesting that PPARδ is associated with GW501516 activity. Furthermore, JNK (c-Jun N-terminal kinase), but not p38 and ERK (extracellular signal-regulated kinase), mediated PPARδ-dependent inhibition of TSP-1 expression in VSMCs exposed to H₂O₂. GW501516- activated PPARδ also reduced the H₂O₂-induced generation of reactive oxygen species, concomitant with inhibition of VSMC migration. In particular, TSP-1 contributed to the action of PPARδ in the regulation of H₂O₂-induced interleukin-1β expression. These results suggest that PPARδ-modulated downregulation of TSP-1 is associated with reduced cellular oxidative stress, thereby inhibiting H₂O₂-induced phenotypic changes in vascular cells.

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Fast LC–MS/MS analysis of free oxysterols derived from reactive oxygen species in human plasma and carotid plaque

Abstract: This rapid LC-MS/MS method enables reliable quantification focused on especially ROS-derived oxysterols in human plasma and atherosclerotic plaque samples under high-throughput conditions.

Cited by 68 publications

References 32 publications

Detection and confirmation of serum lipid biomarkers for preeclampsia using direct infusion mass spectrometry

Detection and confirmation of serum lipid biomarkers for preeclampsia using direct infusion mass spectrometry

Selective changes in cholesterol metabolite levels in plasma of breast cancer patients after tumor removal

Ligand-Activated Peroxisome Proliferator-Activated Receptor δ Attenuates Vascular Oxidative Stress by Inhibiting Thrombospondin-1 Expression

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